Effects of 6-week aerobic exercise on calcium regulation in skeletal muscle of ApoE knockout mice fed by high-fat diet

doi:10.12047/j.cjap.6221.2022.025

Abstract

Abstract: Objective: To investigate the effects of 6 weeks of aerobic exercise on the sarcoplasmic reticulum calcium regulatory proteins in skeletal muscle of apolipoprotein E (ApoE) knockout mice fed by high-fat diet. Methods: There were a total of twenty five 9-week-old ApoE knockout mice (ApoE knockout mice, ApoE KO), five of which were selected randomly for the maximum running speed test. The running speed would be increased by 1.2 m/min every 3 min after a 5-min duration of initial speed of 4.8 m/min without slope until exhaustion, then the final speed was set as maximal speed, and the test result of the maximum running speed was (27.0±2.4)m/min. The remaining 20 ApoE KO mice were randomly divided into ApoE KO mouse high-fat diet group (KO) and ApoE KO mouse high-fat diet + aerobic exercise group (KE), 10 mice per group. Ten 9-week-old wild-type C57BL/6J mice were used as a blank control group (wild-type, WT). High fat diet composition: fat content was 21% (w/w) and cholesterol content was 1.5% (w/w). Exercise intervention was initiated 1 week after adaptive training in the KE group with an exercise protocol consisting of 40% maximal running speed (10.8 m/min), 40 min/d and a frequency of 3 d/w for a total of 6 weeks. All mice were anesthetized and sacrificed by cardiac puncture then bilateral gastrocnemius muscles were isolated immediately 48 h after the final exercise. Skeletal muscle Ca²⁺ concentration was detected by visible light colorimetry; the expression levels of RyR, CaM, CaMK Ⅱ, SERCA1 and SERCA2, which are calcium regulated proteins of sarcoplasmic reticulum, in mouse skeletal muscle, was detected by Western blotting. Results: Compared with WT mice, the Ca²⁺ concentration, the expression levels of the sarcoplasmic reticulum calcium releasing proteins RyR, CaMKⅡ, and the calcium recycling proteins SERCA1 and SERCA2 were decreased significantly in skeletal muscle of the KO mice (P＜0.05, P＜0.01), while the expression of CaM protein did not change. Skeletal muscle Ca²⁺ concentration and the levels of sarcoplasmic reticulum calcium recycling proteins SERCA1 and SERCA2 were increased significantly (P＜0.05) in KE mice compared with KO mice, but there were no significant differences in the expressions of the sarcoplasmic reticulum calcium release proteins RyR, CaM and CaMK II. Conclusion: High fat diet can reduce the concentration of Ca²⁺in skeletal muscle and weaken the release and recovery of sarcoplasmic reticulum calcium in ApoE knockout mice. 6-week aerobic exercise training can significantly increase its Ca²⁺concentration and promote the recovery of sarcoplasmic reticulum calcium.

Key words: apolipoprotein E, aerobic exercise, skeletal muscle, calcium regulation, mice, gene knockout

CLC Number:

Q445

CHEN Jun, GAO Yang, SU Xiang-nan, YU Liang. Effects of 6-week aerobic exercise on calcium regulation in skeletal muscle of ApoE knockout mice fed by high-fat diet[J]. CJAP, 2022, 38(2): 137-142.

References

[1] Kim K, Anderson EM, Scali ST, et al. Skeletal muscle mitochondrial dysfunction and oxidative stress in peripheral arterial disease: A unifying mechanism and therapeutic target[J]. Antioxidants(Basel), 2020, 9(12): E1304-1327.
[2] Gasparini M, Sabovic M, Gregoric ID, et al. Increased fatigability of the gastrocnemius medialis muscle in individuals with intermittent claudication[J]. Eur J Vasc Endovasc Surg, 2012, 44(2): 170-176.
[3] McDermott MM, Ferrucci L, Guralnik J, et al. Pathophysiological changes in calf muscle predict mobility loss at 2-year follow-up in men and women with peripheral arterial disease[J]. Circulation, 2009, 120(12): 1048-1055.
[4] Sfyri P, Matsakas A. Crossroads between peripheral atherosclerosis, western-type diet and skeletal muscle pathophysiology: emphasis on apolipoprotein E deficiency and peripheral arterial disease[J]. J Biomed Sci, 2017, 24(1): 42-67.
[5] 冯利霞, 葛长江, 吕树铮, 等. 细胞钙离子转运蛋白兰尼碱受体3与ApoE基因敲除小鼠动脉粥样硬化斑块形成的关系[J]. 中国病理生理杂志, 2012, 28(9): 1686-1689.
[6] 郑晔. RYR3、MiR-29b及MMP-9在小鼠动脉硬化斑块形成过程中的作用[D]. 天津医科大学, 2017.
[7] Ewart M-A, Kennedy S, Macmillan D, et al. Altered vascular smooth muscle function in the ApoE knockout mouse during the progression of atherosclerosis[J]. Atherosclerosis, 2014, 234(1): 154-161.
[8] Edwards JM, Neeb ZP, Alloosh MA, et al. Exercise training decreases store-operated Ca²⁺ entry associated with metabolic syndrome and coronary atherosclerosis[J]. Cardiovasc Res, 2010, 85(3): 631-640.
[9] Lavier J, Beaumann M, Ménetrey S, et al. Supramaximal intensity hypoxic exercise and vascular function assessment in mice[J]. J Vis Exp, 2019(145): e58708-58717.
[10] Van Assche T, Fransen P, Guns PJ, et al. Altered Ca²⁺ handling of smooth muscle cells in aorta of apolipoprotein E-deficient mice before development of atherosclerotic lesions[J]. Cell Calcium, 2007, 41(3): 295-302.
[11] Munkvik M, Rehn TA, Slettaløkken G, et al. Training effects on skeletal muscle calcium handling in human chronic heart failure[J]. Med Sci Sports Exerc, 2010, 42(5): 847-855.
[12] Denniss A, Dulhunty AF, Beard NA. Ryanodine receptor Ca²⁺ release channel post-translational modification: Central player in cardiac and skeletal muscle disease[J]. Int J Biochem Cell Biol, 2018, 101: 49-53.
[13] Bellinger AM, Reiken S, Dura M, et al. Remodeling of ryanodine receptor complex causes “leaky” channels: a molecular mechanism for decreased exercise capacity[J]. Proc Natl Acad Sci U S A, 2008, 105(6): 2198-2202.
[14] Bueno CR, Ferreira J C B, Pereira MG, et al. Aerobic exercise training improves skeletal muscle function and Ca²⁺handling-related protein expression in sympathetic hyperactivity-induced heart failure[J]. J Appl Physiol(1985), 2010, 109(3): 702-709.
[15] Mänttiri S, Anttila K, Kaakinen M, et al. Effects of low-intensity training on dihydropyridine and ryanodine receptor content in skeletal muscle of mouse[J]. J Physiol Biochem, 2006, 62(4): 293-301.
[16] Fodor J, Gomba-Tóth A, Oláh T, et al. Alteration of sarcoplasmic reticulum Ca²⁺ ATPase expression in lower limb ischemia caused by atherosclerosis obliterans[J]. Physiol Int, 2017, 104(2): 183-192.
[17] 丁海丽, 黄增浩, 任在方, 等. 针刺对大鼠运动性骨骼肌损伤内质网应激的干预作用及机制[J]. 中国应用生理学杂志, 2021, 37(4): 359-364.
[18] Xu H, Van Remmen H. The SarcoEndoplasmic Reticulum Calcium ATPase(SERCA) pump: a potential target for intervention in aging and skeletal muscle pathologies[J]. Skeletal Muscle, 2021, 11(1): 25.
[19] Melo SFS, Barauna VG, Neves VJ, et al. Exercise training restores the cardiac microRNA-1 and -214 levels regulating Ca²⁺ handling after myocardial infarction[J]. BMC Cardiovasc Disord, 2015, 15: 166-174.
[20] Morissette MP, Susser SE, Stammers AN, et al. Exercise-induced increases in the expression and activity of cardiac sarcoplasmic reticulum calcium ATPase 2 is attenuated in AMPKα2 kinase-dead mice[J]. Can J Physiol Pharmacol, 2019, 97(8): 786-795.